Sin1 and mTOR differentially regulate TGF-β dependent Treg differentiation.
a)
Sin1+/+ (WT) or Sin1−/− (KO) CD4+ T cells were stimulated in vitro with anti-CD3 plus anti-CD28 antibodies in the presence or absence of 1 ng/ml TGF-β. After four days Foxp3 expressing cells were enumerated by flow cytometry. Data shown are representative of two independent experiments.
b)
Sin1+/+ CD4+ T cells were stimulated in vitro with anti-CD3 plus anti-CD28 antibodies plus 1 ng/ml TGF-β in the presence of rapamycin (Rapa. 30 nM), pp242 (100 nM) or carrier (ctrl.). The proportion of Foxp3+ cells generated after four days was measured by flow cytometry. Data are representative of two independent experiments.
c)
Sin1+/+ CD4+ T cells were activated with anti-CD3 in the presence of indicated inhibitors and assayed for induction of S6 p-S235/S236 and Akt p-S473 by flow cytometry. Data are representative of two independent experiments.